细菌诱导光滑鳖甲幼虫抑制差减cDNA文库

作者刘忠渊，王鹏，毛新芳摘要，关键词

摘要：本研究以光滑鳖甲幼虫的cDNA为材料，应用抑制差减杂交(SuppressionSubtractiveHybridization，SSH)技术构建细菌诱导的光滑鳖甲幼虫抑制差减cDNA文库，并利用反向Northern杂交技术获得差异基因．结果表明，随机挑取的阳性克隆的大小均为～bp，符合差减文库PCR产物片段的大小.通过斑点杂交技术共筛选出个差异基因片段，对其克隆、测序及同源性分析后鉴定出70种编码蛋白的基因ESTs序列，所得ESTs序列主要涉及免疫防御、代谢过程、生长发育类等相关基因．细菌诱导光滑鳖甲幼虫抑制差减cDNA文库的构建，为进一步克隆光滑鳖甲免疫抗菌相关基因及其免疫防御机制的研究奠定了基础．

关键词:光滑鳖甲；抑制差减杂交；cDNA文库；反向Northern

AbstractKeywords

ConstructionandAnalysisofSuppressionSubtractivecDNALibraryofAnatolicaPolitaBorealisLarvaInducedbyBacteria

LIUZhongyuan,WANGPeng,MAOXinfang

Abstract:Inthepresentstudy,suppressionsubtractivecDNAlibrarywasconstructedforAnatolicapolitaborealislarvainducedbybacteriausingsuppressionsubtractivehybridization(SSH)technique,anddifferentialgeneswereobtainedbyreverseNorthernblot.Theresultsshowedthattherandomlyselectedpositivecloneswereallin-bp,inaccordwiththepredictionforPCR,differentialgenesfragmentswerescreenedbydotblothybridization.Theexpressedsequencetags(ESTs)sequencesof70proteinswereidentifiedbysequenceandhomologyanalysis.TheESTssequencesweremainlyinvolvedinimmunedefense,metabolicprocesses,growingdevelopment,andotherrelatedgenes.ConstructionofsuppressionsubtractivecDNAlibraryofA.politalarvainducedbybacteria,forfurthercloningA.politaimmuneantibacterial.relatedgeneanditsimmunedefensemechanismresearchlaidthefoundation.

Keywords:Anatolicapolitaborealis;SuppressionSubtractiveHybridization;cDNAlibrary;ReverseNorthern

文献来源本文载于新疆大学学报(自然科学版)年第2期文献信息

DOI:10./j.cnki...02.

中图分类号:Q

文献标识码:A

文章编号:-()02--06

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